JP2008054858A - Two liquid type adhesive - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an adhesive composition for living tissue which allows a user to control not only adhesive time but also gelling reaction rate more preferably compared to conventional compositions. <P>SOLUTION: Two-liquid type adhesive comprising a main agent and a cross-linking agent is provided. The main agent contains genetically engineered albumin, and the cross-linking agent contains trehalose aldehyde and glutaraldehyde. When using the two-liquid type adhesive, the main agent and the cross-linking agent are mixed at a weight ratio of 100:1 to 100:40. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a two-component adhesive. More specifically, the present invention relates to a two-component adhesive for living tissue adhesion having hemostasis and bioabsorbability.

  Various surgical tissue adhesives are used. Of these, cyanoacrylates, gelatin-formaldehyde compositions and fibrin-based adhesives have received the most attention.

  Cyanoacrylate adhesives have been studied for surgical use. For example, certain isobutyl cyanoacrylate formulations have been approved for use in livestock treatment. Typically, a monomer and / or mixture of monomers is applied to the adhesion site and rapidly polymerizes into a solid. One drawback of cyanoacrylate adhesives is that the affected area needs to be dry. The other is that the solid obtained by polymerization is not absorbed into the body, and for these reasons, the usefulness of this adhesive is reduced when used in vivo. In addition, polymerization is accompanied by heat generation, and the results of poor tissue compatibility have been reported (see Non-Patent Documents 1 to 3).

  Gelatin-formaldehyde adhesives are based on gelatin cross-linked with formaldehyde and have been used experimentally mainly in Europe since 1964. Of the proposed formulations, GRF (gelatin, resorcin, formaldehyde) is the best known. A heated solution of gelatin is mixed in situ with a hardener consisting of a formaldehyde-based solution, and the mixture quickly solidifies and adheres to the tissue. For GRF adhesives, there is a concern that formaldehyde is essential. Further, gelatin needs to be applied under heating, for example, at a temperature higher than body temperature. Furthermore, the technique of mixing and application has not been strictly determined under the clinical practice of GRF (see Non-Patent Documents 4 to 7).

Fibrin glue utilizes a blood clotting system to produce an adhesive or sealant composition. One commercially available composition is Tussicol® available from Rugis, France. The other is Fibrin Sealant Kit 1.0, available from Osterreichisches Institut for Hasmoderirate, GMBH (Under Immuno GA, A-1220, Vienna, Austria). When fibrin glue is used, blood aggregates are formed by two types of components. One of the components is a solution of fibrinogen and blood clotting factors, such as factor XIII, and the other is a solution of thrombin and calcium ions. The disadvantages of fibrin adhesives are weak tensile strength (generally less than 50 g / cm ² ) and slow cure time. There is also the risk of viral infection in patients when using blood products (fibrinogen and cofactors) from multiple human donors. A method using autologous blood for preparing a fibrin sealant has been proposed (see Patent Document 1).

  An adhesive composition for living tissue is known in which gelatin, which is a protein, is used as a main agent and glutaraldehyde or the like is used as a cross-linking agent (see Patent Document 2). However, glutaraldehyde alone is insufficient to control the gelation reaction time and gelation reaction rate. For example, if the concentration of the crosslinking agent is increased in order to shorten the gelation reaction time, it is solidified immediately after mixing the main agent and the crosslinking agent, and a sufficient adhesive effect cannot be expected. In addition, when the crosslinking agent concentration is lowered, a gentle gelation reaction proceeds, but a complete gel cannot be obtained. This is because protein elution occurs due to moisture such as blood, which causes a problem that long-term adhesive strength cannot be ensured.

  For these reasons, no technology has been established so far that achieves both the adhesion time according to the application by controlling the gelation completion time and maintaining the adhesive force by complete gelation. These techniques are particularly useful in optimizing the composition when the main agent and the crosslinking agent are automatically mixed and applied by the device.

  An adhesive for biological tissue of a genetic manipulation-derived protein and a polyfunctional aldehyde is known (see Patent Document 3). In the examples, only glutaraldehyde is used as the crosslinking agent. In addition, an adhesive for living tissue using a collagen protein partial hydrolyzate and a water-soluble chitin derivative as a main agent and an alkylenedialdehyde as a crosslinking agent is known (see Patent Document 4). In the examples, glutaraldehyde is used as the crosslinking agent. Furthermore, an adhesive sealant composition using a protein within a specific pH range as a main ingredient and a modified polyethylene glycol as a cross-linking agent is known (see Patent Document 5). However, since the usable pH is limited, its versatility is limited.

In addition, an adhesive composition using plasma protein as a main ingredient and a polyfunctional aldehyde as a crosslinking agent is known (see Patent Document 6). In this patent document, examples using aldehydes derived from trehalose are disclosed. However, since trehalose aldehyde alone is used as a cross-linking agent, it takes a long reaction time of 1 hour to cure the adhesive, and the tear strength after 1 hour is only 245 g / cm ² . Not practical as an adhesive.
JP-A-57-149229 Japanese Patent Laid-Open No. 6-70972 JP 2000-288079 A JP 2000-290633 A Japanese Patent No. 3592718 Japanese Patent No. 3483882 Bonutti, P. et al., Clinical Orthopedics and Related Research, Vol. 229, pages 241-248 (1988) Nelson, R. et al., Arch. Surg. Vol. 100, pages 295-298 (1970) Celik, H., et al., “Nonsuture closure of arterial defect by vein graft using isobutyl-2-cyanoacrylates as a tissue adhesive”, Journal of Nearosurgical Sciences Vol. 35, No. 2 (April-June 1991) Fabiani, et al., Ann. Thorac. Surg. 143-145 (1990) Fabiani, et al., Supplement J. Circulation Vol. 80, No. 3 (September 1989) Bachet, et al., J. Thorac. Cardiovasc. Surg. Vol. 83 (1982) Braunwald, et al., Surgery Vol. 59, No. 6 (June 1966)

  It is an object of the present invention to provide a two-component adhesive for living tissue adhesion that is superior not only in controlling the adhesion time but also in controlling the gelation reaction rate as compared with the prior art.

The inventors of the present invention not only can control the adhesion rate by using a mixture of a crosslinking agent composed of trehalose aldehyde and glutaraldehyde and a main component composed of genetically modified albumin, but also a gelation reaction that is important for the durability of living tissue adhesion. The inventors have found that the rate can be controlled and have completed the present invention. That is, the present invention has the following configuration.
[1] A two-component adhesive comprising a main agent and a cross-linking agent, wherein the main agent contains recombinant albumin, the cross-linking agent contains trehalose aldehyde and glutaraldehyde, and the main agent and the cross-linking agent are in a weight ratio of 100: A two-component adhesive for use in a mixture of 1 to 100: 40.
[2] The two-component adhesive according to [1], wherein the total content of trehalose aldehyde and glutaraldehyde in the crosslinking agent is 2 to 45% by weight.
[3] The two-component adhesive according to [1] or [2], wherein the content of the recombinant albumin in the main agent is 10 to 55% by weight.
[4] The two-component adhesive according to any one of [1] to [3], wherein the trehalose aldehyde content in the crosslinking agent is 1 to 30% by weight.
[5] The two-component adhesive according to any one of [1] to [4], wherein the content of glutaraldehyde in the crosslinking agent is 1 to 15% by weight.
[6] The two-component adhesive according to any one of [1] to [5], wherein trehalose aldehyde and glutaraldehyde in the crosslinking agent are in a ratio of 20: 1 to 1: 2 by weight.
[7] The two-component adhesive according to any one of [1] to [6], which is an adhesive for living tissue.
[8] The two-component adhesive according to any one of [1] to [7] and the adhesive can be used or used for tissue adhesion, closure, or prevention of blood or body fluid leakage during surgery. A commercial package containing a statement about the adhesive stating that it should be.

  The two-component adhesive of the present invention is a ready-to-use adhesive that not only controls the adhesion time but also controls the gelation reaction rate better than conventional adhesives. The control of the adhesion time in the present invention means that the gelation completion time can be controlled according to the use of the two-component adhesive, and according to the present invention, the two-component that enables the precise mixing of the main agent and the crosslinking agent. It is possible to provide a material useful for optimizing the bonding time when bonding a living tissue using a mixing device or the like. Moreover, since the adhesive agent of this invention has favorable control of a gelation reaction rate, it can provide the material which can improve adhesive durability.

  The two-component adhesive of the present invention is composed of a main agent and a crosslinking agent.

  The main agent contains recombinant albumin as an adhesive component. The genetically modified albumin in the present invention refers to albumin produced by a genetic engineering technique and excluding naturally occurring albumin. Preferably, genetically modified high-purity human serum albumin (γHSA) obtained by highly purifying albumin obtained by genetic engineering techniques is used. By using such genetically modified albumin, unlike natural albumin purified from blood, it is possible to provide an adhesive that is free from animal or human-derived virus contamination and is excellent in safety and quality stability.

  The recombinant albumin and γHSA can be produced by a method known per se. That is, a recombinant albumin or γHSA producing host is obtained by a conventional method, and the target recombinant albumin or γHSA is produced by culturing it using a known culture method, and then the host-related components or other contaminating components are removed. To do. In order to suppress coloring, it is preferable to subject the recombinant albumin or γHSA to a further sufficient purification step.

  A general property of γHSA preferably used in the present invention is a single substance having a molecular weight of about 67,000, a polymer including a dimer, or a decomposition product (molecular weight of about 43,000). Is substantially not included. Furthermore, γHSA is substantially free of contaminating components or polysaccharides derived from the production host, particularly contaminating components or polysaccharides having antigenicity.

  The content of the genetically modified albumin in the main agent is preferably 10 to 55% by weight, more preferably 16 to 50% by weight, still more preferably 22 to 47% by weight, particularly preferably 27 to 47% by weight. %. Within such a range, a sufficient adhesive strength can be obtained, and the viscosity of the main agent is suitable and easy to handle, which is preferable.

The main ingredient contains a solvent in addition to the adhesive component, and may contain a pharmaceutically acceptable additive as desired. Examples of the solvent include water, N-methyl-2-pyrrolidone and the like, preferably distilled water and distilled water for injection.
Examples of the additive include N-acetyltryptophan sodium, sodium caprylate, and sodium bicarbonate, which are albumin stabilizers. The amount of the additive added is preferably 0.00001 to 0.1% by weight in the present invention although it depends on the albumin concentration.

  The crosslinking agent in the present invention contains trehalose aldehyde (TA) and glutaraldehyde (GA) as crosslinking components. The crosslinking component has a function of polymerizing and gelling genetically modified albumin which is an adhesive component in the main agent.

  In the present invention, gelation means that by mixing the main agent and the crosslinking agent, a polymerization reaction occurs, the molecular weight of the adhesive component becomes infinite, and a swollen body insoluble in any solvent is formed. The reaction accompanying the gelation is called a gelation reaction.

  In medicine and biology, aqueous aldehyde solutions are widely used for preserving and fixing treatments for biological tissue specimens. This is because the aldehyde penetrates into and out of the tissue and binds mainly to the protein in the tissue, thereby damaging the three-dimensional structure of the protein and inhibiting various biological activities such as enzyme activity, transport and secretion. It is thought to be because. Since ancient times, aldehydes have been used to immobilize biological tissues, and glutaraldehyde treatment has been used as a preparation stage for specimen preparation for microscopic observation. Alkylenedialdehyde represented by glutaraldehyde has a very high reaction rate. The use of alkylene dialdehyde is indispensable in order to show a high reactivity in an aqueous solution and to obtain a bonding time on the order of minutes.

  Formaldehyde and glutaraldehyde are used as the above-mentioned cross-linking agent for GRF, which is an adhesive for living tissues, but recently there is a concern about their biotoxicity. This is because the aldehyde group of alkylenedialdehyde reacts with proteins such as enzymes that make up the living body and may introduce hydrophobic alkylene groups into the molecule, thereby losing enzyme activity and the like. If it remains, there is a risk of causing toxicity that inhibits life activity as described above. That is, when using a tissue adhesive together with a surgical suture, it is preferable that the gel time is as short as possible from the viewpoint of shortening the operation time and reducing the burden on the patient. There is a problem in terms of biological safety of the reactant. Therefore, as a result of intensive investigations on improving the safety of adhesives for biological tissues, the present inventors have found that unreacted substances remain in the living body if saccharides essential for human life activities can be aldehyded and used. Even if it reacts with protein molecules, we thought that the effect of sugar can be minimized by the hydrophilic structure of sugar, so we focused on sugar aldehyde.

  In the gelation reaction between the main agent and the crosslinking agent, when GA is used alone as the crosslinking agent, the gelation reaction proceeds rapidly and the gelation reaction proceeds rapidly. However, there is a concern about toxicity when the crosslinking agent remains. On the other hand, when TA alone is used as a cross-linking agent, the safety to the living body is enhanced, but the reactivity is low compared to GA alone, and it is difficult to complete the gelation reaction early after mixing the two liquids. Then, it discovered that it could harden | cure in the controlled reaction time, suppressing residual of an unreacted substance by mixing and using highly safe TA and highly reactive GA.

  In the present invention, the sugar chain part of the sugar aldehyde preferably has 1 to 1000 repeating units. More preferably, it is 1-100, More preferably, it is 1-10, Most preferably, it is 1-2. Specifically, glucose, galactose, mannose, fucose, xylose, N-acetylglucosamine, N-acetylgalactosamine, N-acetylneuramin, sucrose, lactose, maltose, isomaltose, cellobiose, trehalose, etc. can give. If the repeating unit is 1000 or less, the gelation reaction proceeds efficiently. The disaccharide trehalose has a function of protecting cells and intracellular substances when frozen or dried, and is a substance that is expected to play a role as a storage and stabilizer in the fields of medicine, cosmetics, food, etc. Therefore, it is preferable as a raw material. Hereinafter, trehalose aldehyde (TA) is used in the present invention.

  Although the preparation method of TA is not specifically limited, The method of oxidatively decomposing saccharides is mentioned as an example. Examples of the oxidizing agent used for oxidative decomposition include chlorine, hypochlorous acid, hydrogen peroxide, potassium permanganate, ammonium persulfate, periodic acid, nitric acid, ozone, etc. Periodic acid or a salt thereof is preferred from the viewpoint that an aldehyde can be produced. Examples of periodic acid salts include sodium periodate and calcium periodate.

  The TA content in the crosslinking agent is preferably 1 to 30% by weight, more preferably 5 to 28% by weight, still more preferably 7 to 26% by weight, and particularly preferably 9 to 23% by weight. Within such a range, the gelation reaction proceeds sufficiently and the toxicity of the aldehyde is not exhibited, which is preferable.

  The GA content in the crosslinking agent is preferably 1 to 15% by weight, more preferably 1 to 12% by weight, still more preferably 2 to 10% by weight, and particularly preferably 3 to 6% by weight. Within this range, the gelation completion time is appropriate and the gelation reaction rate is sufficient. Also, the risk of toxicity due to residual aldehyde is reduced.

  In the present invention, as described above, TA and GA are mixed at a specific ratio as a cross-linking agent in order to achieve a high level of contradictory properties such as safety for a living body and shortening of gelation completion time and consequently shortening of operation time. And preferably used. The total content of TA and GA contained in the cross-linking agent is preferably 2 to 45% by weight, more preferably 5 to 30% by weight, and 7 to 30% by weight from the viewpoint of achieving both safety and reactivity. Even more preferred is 12 to 28% by weight.

  The ratio of TA and GA in the crosslinking agent is preferably 20: 1 to 1: 2 by weight. If the TA concentration is too low, the gelation may be completed too early and the gelation may be completed before application, and the adhesive ability may not be exhibited. Further, if the TA concentration in the cross-linking agent is too high, the completion of gelation is too late, which requires extra adhesion time and may cause a problem that the operation time is delayed. From the above viewpoint, the ratio of TA to GA is a weight ratio, more preferably 20: 1 to 2: 3, still more preferably 20: 1 to 1: 1, particularly preferably 10: 1 to 1: 1. It is.

  In addition to the crosslinking components (TA and GA), the crosslinking agent contains a solvent and may optionally contain a pharmaceutically acceptable carrier. Examples of the solvent include water and ethanol, preferably distilled water and distilled water for injection.

  The two-component adhesive of the present invention is characterized in that the main agent and the crosslinking agent are mixed and used at a weight ratio of 100: 1 to 100: 40 immediately before use. From the viewpoint of reducing the adhesive strength contributed by the crosslinking agent and the possibility that the crosslinking agent GA remains, the mixing ratio is preferably 100: 3 to 100: 35, and more preferably 100: 5 to 100: 30. If it is in the range of this mixing ratio, the adhesive agent excellent in control of adhesion time (gelation reaction time) and gelation reaction rate will be formed.

  In the present invention, the gelation completion time is used synonymously with the adhesion time, and is preferably 30 seconds or more and less than 120 seconds. 35 seconds or more and less than 110 seconds are more preferable, and 40 seconds or more and less than 100 seconds are more preferable. If the gelation completion time is too early, gelation proceeds before application to the application site, and thus the adhesive force may be lost. If the gelation completion time is too late, it takes a long time to bond, and the operation time may be delayed. The gelation completion time is a value measured by the method described in the examples.

  In the present invention, the gelation reaction rate is preferably 80% by weight or more. 82 weight% or more is more preferable and 85 weight% or more is further more preferable. If the gelation reaction rate is too low, a problem of peeling of the bonded portion occurs as a result of elution due to moisture after bonding. The gelation reaction rate is a value measured by the method described in Examples.

  Since the two-component adhesive of the present invention contains albumin as a constituent component, it is preferably used as an adhesive for living tissue. When used for living tissue, it is absorbed by the living body after a certain period of time. When used for living tissue, it is preferably used for tissue adhesion, closure or prevention of blood or body fluid leakage during surgery.

  As an example, a method for applying the two-component adhesive of the present invention to a living tissue will be described. After adding a sufficient amount of the main agent to the living tissue to be applied, the application site can be adhered by adding only the minimum necessary amount of the cross-linking agent, immediately after mixing uniformly, and waiting until gelation is completed.

  The present invention describes the two-component adhesive of the present invention and that the adhesive can or should be used for tissue adhesion, closure or prevention of blood or body fluid leakage during surgery. A commercial package is provided that includes a description of the adhesive.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited by these.

(Measurement of gel completion time)
This was carried out using a dynamic viscoelasticity measuring machine (RS1 manufactured by HAAKE). A disposable parallel plate sensor system was used under the conditions of stress: 100 Pa, frequency: 1 Hz, measurement temperature: 20 ° C., gap: 1 mm. The measurement sample was prepared as follows. The main agent was added on the parallel plate, and the crosslinking agent was added in an amount shown in the Examples. The measurement was started immediately after mixing it with a spatula for 5 seconds. The gelation time was evaluated by the position where the elastic term (G ′) and the viscous term (G ″) intersect (G ′ = G ″), that is, the gel point. A gel point of 40 seconds or more and less than 100 seconds was considered good, and less than 40 seconds and 100 seconds or more were considered bad.

(Measurement of gelation reaction rate)
In a 20 mL glass vial, the main agent and the crosslinking agent were added in the amounts shown in the Examples, and the mixing of both was continued with a spatula for 10 minutes. 20 mL of physiological saline was added to the gel obtained by mixing, and incubation was performed at 37 ° C. with shaking for 24 hours. The gel after the incubation was taken out and frozen at −5 ° C. for 24 hours, and freeze-dried for 48 hours in a vacuum dryer. The gelation reaction rate was calculated by dividing the dry weight value after incubation by the sum of the theoretical dry weight value of the main agent and the crosslinking agent before mixing. When the gelation reaction rate was 80% by weight or more, it was considered good, and when it was less than 80% by weight, it was regarded as poor.

(Tensile strength measurement)
The main agent and then the crosslinking agent were added in a plastic petri dish with a diameter of 60 mm, and after mixing them for 5 seconds with a spatula, the end of a wool specimen (0.3 × 10 × 50 mm) 10 × 10 mm After coating within the range and adhering to the tip of another test piece, it was pressed with a 50 g weight for 2 minutes. Thereafter, evaluation by a tensile tester was performed. The tensile tester was an Autograph AGS-50NJ (Shimadzu Corporation). The test conditions are as follows. The load cell was 5 kg, the crosshead speed was 30 mm / min, and the distance between chucks was 7 cm. Average values (n = 5) were used as data.

(Cytotoxicity test)
Ministry of Health, Labor and Welfare administrative contact Medical Device Examination No.36 ("Reference Materials on Basic Concepts of Biological Safety Tests" March 19, 2003) and ISO 10993-5 (Test for In Vitro Cytotoxicity, May 15, 1999) and was carried out according to the following procedure.
An extract obtained by performing extraction at 37 ° C. for 24 hours in a medium at a rate of 10 ml per 1 g of the gelled product after the reaction between the main agent and the crosslinking agent was used as a test solution. The test solution was diluted in various concentration ranges, and V79 cells were cultured at each concentration for 6 days in a 37 ° C.-CO ₂ incubator. From the culture results, the colony formation rate of each test group with respect to a negative control containing only the medium was determined, and the concentration (IC ₅₀ value) that inhibits the colony formation rate by 50% was determined.

(Production Example 1) Production of crosslinking agent (synthesis of trehalose aldehyde)
Α, α-trehalose dihydrate (endotoxin-free) (Hayashibara) dried under reduced pressure for 2 hours at 60 ° C. (using a water bath manufactured by Yazawa Seisakusho) at 2 mmHg (using a vacuum pump GDH-60 manufactured by Shimadzu Corp.) 19.27 g was dissolved in 200 mL of ion exchange water (Kishida Chemical) and charged into a 2 L four-necked flask. Periodic acid dihydrate (Aldrich) (48.75 g) was dissolved in ion-exchanged water (Kishida Chemical) (400 mL) and charged into the four-necked flask. 200 mL of ion-exchanged water was charged into the four-necked flask. The four-necked flask was stirred for 22 hours at 21 ° C. (using a low temperature and high temperature tank CA-1111 manufactured by Tokyo Science Instruments) while stirring under light shielding (using a stirrer PC-30 manufactured by Nonaka Science Instruments). 1 L of ion exchange resin (AmberliteIRA96SB: organo) was washed with 500 mL of ion exchange water three times. The reaction solution was transferred to a 2 L beaker, and an ion exchange resin was added while stirring (using a magnetic stirrer SRS710AA manufactured by Advantech). Ion exchange resin 600 mL was added, and it was confirmed that the pH of the aqueous layer (Advantech test paper UNIV) was neutral. The ion exchange resin was separated by filtration (Advantech filter paper No. 2) and washed four times with 400 mL of ion exchange water. The filtrate and the washing solution were transferred to an eggplant-shaped flask, and concentrated and dried under reduced pressure at 2 mmHg at 35-40 ° C. (using a water bath manufactured by Yazawa Seisakusho) to obtain 16.61 g of trehalose aldehyde (TA).

(Adjustment of crosslinker concentration)
The concentration was adjusted by dissolving TA in distilled water for injection (Otsuka Pharmaceutical Co., Ltd.). The concentration of glutaraldehyde (GA) was adjusted by diluting a commercially available 25% by weight solution with distilled water for injection. When two types of cross-linking agents are mixed and used, for example, the expression “10% TA + 4% GA” means that 10 wt% TA and 4 wt% GA coexist in the cross-linking agent.

(Production Example 2) Production of main agent γHSA 25% aqueous solution (GMO Albumin, Bifa Co., Ltd.) was lyophilized to obtain γHSA powder, and a predetermined amount was dissolved in distilled water for injection (Otsuka Pharmaceutical Co., Ltd.) to produce γHSA A main agent consisting of an aqueous solution was prepared.

(Example 1)
Various measurements were performed using 0.5 g of the main agent of Production Example 2 and 0.05 g of a 21% TA + 4% GA cross-linking agent.

(Example 2)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.05 g of 10% TA + 5% GA crosslinking agent.

(Example 3)
Various measurements were performed using 0.5 g of the main agent of Production Example 2 and 0.05 g of a 21% TA + 4% GA cross-linking agent.

(Comparative Example 1)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.25 g of a 21% TA + 4% GA crosslinking agent.

(Comparative Example 2)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.0025 g of 21% TA + 4% GA crosslinking agent.

(Comparative Example 3)
Various measurements were performed using 0.5 g of the main agent of Production Example 2 and 0.25 g of a 5% GA crosslinking agent.

(Comparative Example 4)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.25 g of a 2.5% GA crosslinking agent.

(Comparative Example 5)
Various measurements were performed using 0.5 g of the main agent of Production Example 2 and 0.0025 g of a 10% GA crosslinking agent.

(Comparative Example 6)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.0025 g of a 2.5% GA crosslinking agent.

(Comparative Example 7)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.25 g of a 24.5% TA + 0.5% GA crosslinking agent.

(Comparative Example 8)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.25 g of a 6% TA + 19% GA crosslinking agent.

(Comparative Example 9)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.0025 g of a 24.5% TA + 0.5% GA crosslinking agent.

(Comparative Example 10)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.0025 g of a cross-linking agent of 6% TA + 19% GA.

(Comparative Example 11)
Various measurements were carried out using 0.5 g of the main agent of Production Example 2 and 0.05 g of a 10% GA crosslinking agent.

  The results are shown in Table 1. As a result of measuring the gelation completion time and the gelation reaction rate, it was possible to control the gelation completion time while maintaining a good gelation reaction rate in the examples. Although the gelation reaction rate is high, there is a problem that the gelation completion time is too early. When GA is low in concentration, the gelation completion time is suitable but the gelation reaction rate is low. Further, the adhesives of the examples had sufficient tensile strength and no problem with cytotoxicity.

  In the present invention, by using a cross-linking agent in which TA and GA are combined in a predetermined ratio with the main agent, gelation is completed in a very short time, and a composition for bonding a living tissue having a better gelation reaction rate is obtained. I found out that

  The two-component adhesive of the present invention can provide a material that is superior not only in controlling the adhesion time but also in controlling the gelation reaction rate as compared with conventional adhesives. The control of the adhesion time means that the gelation rate can be controlled according to the use of the adhesive. Furthermore, it is possible to provide a material that is particularly useful for optimizing the adhesion time during biological tissue adhesion using a two-component mixing device that enables precise mixing of the main agent and the crosslinking agent. Moreover, the material which can improve adhesion durability can be provided by control of a gelation reaction rate.

Claims (8)

  A two-component adhesive comprising a main agent and a cross-linking agent, the main agent containing genetically modified albumin, the cross-linking agent containing trehalose aldehyde and glutaraldehyde, and the main agent and the cross-linking agent in a weight ratio of 100: 1 to 100 : A two-component adhesive for use by mixing at a ratio of 40.   The two-component adhesive according to claim 1, wherein the total content of trehalose aldehyde and glutaraldehyde in the crosslinking agent is 2 to 45% by weight.   The two-component adhesive according to claim 1 or 2, wherein the content of the genetically modified albumin in the main agent is 10 to 55% by weight.   The two-component adhesive according to any one of claims 1 to 3, wherein a trehalose aldehyde content in the crosslinking agent is 1 to 30% by weight.   The two-component adhesive according to any one of claims 1 to 4, wherein the content of glutaraldehyde in the crosslinking agent is 1 to 15% by weight.   The two-component adhesive according to any one of claims 1 to 5, wherein trehalose aldehyde and glutaraldehyde in the cross-linking agent are in a weight ratio of 20: 1 to 1: 2.   The two-component adhesive according to any one of claims 1 to 6, which is an adhesive for living tissue.   The two-component adhesive according to any one of claims 1 to 7, and the fact that the adhesive can or should be used for tissue adhesion, closure or blood or body fluid leakage prevention during surgery. A commercial package containing a description of the adhesive.